LOCUS       sequence-182696-        4473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry TOPO TA cloning vector ERF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4473)
  AUTHORS   Bouveret R, Waardenberg AJ, Schonrock N, Ramialison M, Doan T, de 
            Jong D, Bondue A, Kaur G, Mohamed S, Fonoudi H, Chen CM, Wouters MA,
            Bhattacharya S, Plachta N, Dunwoodie SL, Chapman G, Blanpain C, 
            Harvey RP
  TITLE     NKX2-5 mutations causative for congenital heart disease retain 
            functionality and are directed to hundreds of targets.
  JOURNAL   Elife. 2015 Jul 6;4. doi: 10.7554/eLife.06942.
  PUBMED    26146939
REFERENCE   2  (bases 1 to 4473)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2015-07-6"; volume: "4. doi"; pages: " 10.7554/eLife.06942"
COMMENT     Sequence Label: pCR8/GW/TOPO-ERF
FEATURES             Location/Qualifiers
     source          1..4473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1816..1915)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1932..1951)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1933..1951)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1956..1972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1956..1972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2403..3194
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3287..3875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3776..3795
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4029..4046
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4205..4232)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4324..4410)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4459..4473,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4473,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctccgaattc gcccttacca tgaagacccc ggcggacaca gggtttgcct
      181 tcccagattg ggcctacaaa ccggagtcat cccctggctc caggcagatc cagctgtggc
      241 actttatcct ggagctgctt cggaaagagg agtaccaggg cgtcatcgct tggcaggggg
      301 actacgggga gtttgtcatc aaggaccctg atgaagtggc tcgcctctgg ggggtccgca
      361 agtgcaaacc ccagatgaac tatgacaagc tgagccgggc tttgcgctat tattacaaca
      421 agcgcattct acacaagacc aaggggaaac ggttcaccta caagttcaac ttcaacaaac
      481 tggtgctggt caattaccct ttcatcgata tggggctggc tgggggtgca gttccccaaa
      541 gcgccccacc agtgccatca ggcggcagcc atttccgctt ccctccctca acaccctctg
      601 aggtgctgtc ccccactgag gatccccgat ctccaccggc ttgttcttca tcatcctctt
      661 ctctcttctc tgctgtggtt gcccgacgcc tgggccgagg ctcagtcagt gactgtagtg
      721 atggcacctc agagctggag gagcctctgg gagaggaccc cagggcacga ccacctggcc
      781 ctccggagct gggtgccttc cgagggcccc ccctggcccg cctcccgcat gaccctggtg
      841 tcttccgtgt ctatcctcgg ccccggggtg gtcctgaacc cctgagtccc ttccctgtgt
      901 cacctttggc tgggcctggc tcccttctac cccctcagct ctccccagct ctgcccatga
      961 ctcccaccca cctggcctac acaccctcac ccacgctgag tcctatgtac cccagtggtg
     1021 gtgggggccc tagtggctca gggggaggtt cccacttctc cttcagtcct gaggacatga
     1081 aacggtacct gcaggcccac acccaaagcg tctacaacta ccacctcagt ccccgcgcct
     1141 tcctgcacta cccagggctg gtggtgcccc agcctcagcg ccctgacaag tgcccactgc
     1201 cgcccatggc accggagacc ccgccggtcc cctcctcagc ctcgtcttcc tcttcctcct
     1261 cttcatcccc gttcaagttt aagctgcagc cacccccgct aggacgccgg cagcgggcag
     1321 ctggagagaa ggctctagga ggcactgaca agagcagtgg tggcagtggc tcgggtggac
     1381 tggctgaggg ggcaggtgca gtagctcccc caccgccacc accccagatt aaggtggagc
     1441 ccatctcaga aggagagtcg gaggaggtgg aggtgactga catcagtgac gaagatgagg
     1501 aagatgggga ggtgttcaag actccccgtg ccccgcctgc accccccaag ccagagcccg
     1561 gagaggcacc gggggtggcc cagtgcatgc cccttaaact gcgctttaag cggcgctgga
     1621 gtgaagactg tcgcctggag gggggcgggt gcctgtctgg gggccctgaa gatgagggtg
     1681 aggacaagaa ggtgcgtggg gacgtgggcc ctggggagtc tgggggaccc cttaccccac
     1741 gacgggtgag ctctgacctc cagcacgcca cagcccaact ctccctggag caccgagatt
     1801 ccaagggcga attcgaccca gctttcttgt acaaagttgg cattataaaa aataattgct
     1861 catcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     1921 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     1981 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc
     2041 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
     2101 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     2161 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     2221 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     2281 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     2341 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     2401 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     2461 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     2521 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     2581 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     2641 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     2701 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     2761 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     2821 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     2881 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     2941 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     3001 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     3061 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     3121 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     3181 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     3241 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3301 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3361 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3421 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3481 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3541 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3601 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3661 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3721 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3781 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3841 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3901 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3961 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4021 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     4081 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4141 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     4201 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     4261 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     4321 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     4381 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
     4441 cgctagcatg gatgttttcc cagtcacgac gtt
//
